Casein active single peptide as well as preparation method and application thereof
A casein and active technology, which is applied in the field of biologically active single peptides capable of promoting fermentation of yogurt and inhibiting post-acidification of yogurt, can solve the problems of heavy sourness, shortened shelf life, and affect the quality of yogurt, and achieve the effect of shortening fermentation time.
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Embodiment 1
[0026] 1. Controllable enzymolysis: Add papain to enzymolyze casein under the conditions of 50°C and pH 6.0 to obtain an enzymolysis solution.
[0027] 2. Enzyme inactivation with enzymatic hydrolysis solution: heat at 90°C for 15 minutes to inactivate the enzyme and terminate the enzymatic hydrolysis reaction.
[0028] 3. Centrifuge the enzyme-inactivated hydrolyzate (6000 x g, 4°C) for 20 minutes, and ultrafilter the supernatant (take the
[0029] After 3000 Dalton components), the macroporous resin NKA-II was used to collect the 10% (w / w) ethanol eluate, followed by Sephadex G-25, the mobile phase was deionized water, and the detection wavelength was 220nm. Collected in order of absorption peaks, such as figure 1 shown. The components with strong activity separated by Sephadex G-25 were subjected to C 18 Separation and purification by high performance liquid chromatography, the separation conditions are: 5% (w / w) acetonitrile and 95% (w / w) deionized water gradient eluti...
Embodiment 2
[0031] 1. Controllable enzymatic hydrolysis: Add papain to enzymolyze casein under the conditions of 55°C and pH 7.5 to obtain an enzymatic hydrolysis solution.
[0032] 2. Enzyme hydrolysis solution inactivation: heat at 95°C for 20 minutes to inactivate the enzyme and terminate the enzymatic hydrolysis reaction.
[0033] 3. Centrifuge the enzyme-inactivated hydrolyzate (6000 x g, 4°C) for 20 minutes, and ultrafilter the supernatant (take the
[0034] After 3000 Dalton components), the macroporous resin NKA-II was used to collect 20% (w / w) ethanol eluate, followed by Sephadex G-25, the mobile phase was deionized water, and the detection wavelength was 220nm. Collected in order of absorption peaks, such as figure 1 shown. The components with strong activity separated by Sephadex G-25 were subjected to C 18 Separation and purification by high performance liquid chromatography, the separation conditions are: 40% (w / w) acetonitrile and 60% (w / w) deionized water gradient eluti...
Embodiment 3
[0036] Before yogurt fermentation, add 50ppm (w / w) of the bioactive peptide HAQQKE obtained in Example 1 to replace milk powder with equal protein, and measure the yogurt fermentation cycle, namely T pH4.5 , the time required for yogurt to ferment to pH4.5, in hours. like image 3 As shown, the yoghurt fermentation period was shortened by 23.00% after adding NPSKENL.
[0037] Example 3: Effects of Bioactive Peptides on Changes in Acidity of Yogurt During Storage Period
[0038] Measure according to the titration acidity of the fermented yoghurt of embodiment 2 during 28 days of storage, i.e. the degree of cleanliness T 0 Variety.
[0039] Test method: Use a 5ml straw to take 5ml of fermented milk that has been stirred evenly, add it to a conical flask containing 25ml of distilled water, add 3 to 4 drops of 1% phenolphthalein indicator, and titrate with 0.1NNaOH to light red, and keep it for 20s without fading. The milliliter of consumed NaOH solution multiplied by 20 is t...
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